CA2524928A1 - Assay method and apparatus - Google Patents

Assay method and apparatus Download PDF

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Publication number
CA2524928A1
CA2524928A1 CA002524928A CA2524928A CA2524928A1 CA 2524928 A1 CA2524928 A1 CA 2524928A1 CA 002524928 A CA002524928 A CA 002524928A CA 2524928 A CA2524928 A CA 2524928A CA 2524928 A1 CA2524928 A1 CA 2524928A1
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Prior art keywords
analyte
specific binding
receptacle
binding agent
detecting
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CA002524928A
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French (fr)
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CA2524928C (en
Inventor
Jeffrey D. Mcbride
Francis Guy Gabriel
Peter J. Delves
John L. A. Fordham
Ian A. Cree
Keith Rawson
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Nalia Systems Ltd
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Individual
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D67/00—Processes specially adapted for manufacturing semi-permeable membranes for separation processes or apparatus
    • B01D67/0081—After-treatment of organic or inorganic membranes
    • B01D67/0088—Physical treatment with compounds, e.g. swelling, coating or impregnation
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
    • B01D61/14—Ultrafiltration; Microfiltration
    • B01D61/18—Apparatus therefor
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D67/00—Processes specially adapted for manufacturing semi-permeable membranes for separation processes or apparatus
    • B01D67/0081—After-treatment of organic or inorganic membranes
    • B01D67/0093—Chemical modification
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D69/00—Semi-permeable membranes for separation processes or apparatus characterised by their form, structure or properties; Manufacturing processes specially adapted therefor
    • B01D69/14—Dynamic membranes
    • B01D69/141—Heterogeneous membranes, e.g. containing dispersed material; Mixed matrix membranes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D69/00—Semi-permeable membranes for separation processes or apparatus characterised by their form, structure or properties; Manufacturing processes specially adapted therefor
    • B01D69/14—Dynamic membranes
    • B01D69/141—Heterogeneous membranes, e.g. containing dispersed material; Mixed matrix membranes
    • B01D69/1411—Heterogeneous membranes, e.g. containing dispersed material; Mixed matrix membranes containing dispersed material in a continuous matrix
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    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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    • B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J19/0046—Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5025—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures for parallel transport of multiple samples
    • B01L3/50255—Multi-well filtration
    • G—PHYSICS
    • G01—MEASURING; TESTING
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    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/5302—Apparatus specially adapted for immunological test procedures
    • G01N33/5304—Reaction vessels, e.g. agglutination plates
    • G—PHYSICS
    • G01—MEASURING; TESTING
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    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
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    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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    • G01N33/54366—Apparatus specially adapted for solid-phase testing
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    • B01J2219/00277—Apparatus
    • B01J2219/00279—Features relating to reactor vessels
    • B01J2219/00306—Reactor vessels in a multiple arrangement
    • B01J2219/00313—Reactor vessels in a multiple arrangement the reactor vessels being formed by arrays of wells in blocks
    • B01J2219/00315—Microtiter plates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00351—Means for dispensing and evacuation of reagents
    • B01J2219/00423—Means for dispensing and evacuation of reagents using filtration, e.g. through porous frits
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
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    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00603—Making arrays on substantially continuous surfaces
    • B01J2219/00639—Making arrays on substantially continuous surfaces the compounds being trapped in or bound to a porous medium
    • B01J2219/00641—Making arrays on substantially continuous surfaces the compounds being trapped in or bound to a porous medium the porous medium being continuous, e.g. porous oxide substrates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
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    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00603—Making arrays on substantially continuous surfaces
    • B01J2219/00659—Two-dimensional arrays
    • B01J2219/00662—Two-dimensional arrays within two-dimensional arrays
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
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    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B01J2219/00722—Nucleotides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
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    • B01J2219/0072—Organic compounds
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B01L2300/0636—Integrated biosensor, microarrays
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0829—Multi-well plates; Microtitration plates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
    • B01L2400/049—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics vacuum
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6816—Hybridisation assays characterised by the detection means
    • C12Q1/6825—Nucleic acid detection involving sensors
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/04—Libraries containing only organic compounds
    • C40B40/06—Libraries containing nucleotides or polynucleotides, or derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
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    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/04—Libraries containing only organic compounds
    • C40B40/10—Libraries containing peptides or polypeptides, or derivatives thereof
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    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
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    • C40B60/14—Apparatus specially adapted for use in combinatorial chemistry or with libraries for creating libraries
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/81—Packaged device or kit
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
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    • Y10S436/00—Chemistry: analytical and immunological testing
    • Y10S436/805—Optical property
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S436/00—Chemistry: analytical and immunological testing
    • Y10S436/807—Apparatus included in process claim, e.g. physical support structures
    • Y10S436/81—Tube, bottle, or dipstick

Abstract

The present invention relates to an apparatus for use in an assay comprising:
(a) at least one first receptacle comprising a fluid inlet and a fluid outlet, (b) a porous membrane, and (c) at least one analyte-specific binding agent, characterised in that said at least one analyte-specific binding agent is immobilised on the underside of said porous membrane relative to the fluid inlet; and to methods of performing said assay.

Claims (104)

1. An apparatus for use in an assay comprising:
(a) at least one first receptacle comprising a fluid inlet and a fluid outlet;
(b) a porous membrane; and (c) at least one analyte-specific binding agent, characterised in that said at least one analyte-specific binding agent is immobilised on the underside of said porous membrane relative to the fluid inlet.
2. An apparatus according to claim 1 wherein the porous membrane is positioned at the fluid outlet.
3. An apparatus according to claim 1 or 2 comprising a plurality of first receptacles in the form of a multi-well plate.
4. An apparatus according to claim 3 in the form of a 96-well plate.
5. An apparatus according to any preceding claim wherein a plurality of analyte-specific binding agents is immobilised on said porous membrane in the form of an array.
6. An apparatus according to claim 5 wherein said plurality of analyte-specific binding agents is immobilised in the form of a micro-array.
7. An apparatus according to claim 5 or claim 6 wherein the array comprises at least one control probe.
8. An apparatus according to any preceding claim wherein said at least one analyte-specific binding agent is a polynucleotide probe.
9. An apparatus according to claim 8 wherein said at least one analyte-specific binding agent is an Expressed Sequence Tag (EST).
10. An apparatus according to any one of claims 1-7 wherein said at least one analyte-specific binding agent is a polypeptide binding agent.
11. An apparatus according to claim 10 wherein said at least one analyte-specific binding agent is an antibody or a fragment thereof.
12. An apparatus according to any preceding claim wherein said at least one analyte-specific binding agent is immobilised directly or indirectly on said porous membrane.
13. An apparatus according to any preceding claim wherein the or each first receptacle is positioned at least in part in a second receptacle which is capable of collecting liquid from the fluid outlet of the first receptacle such its porous membrane may be submerged in said liquid.
14. An apparatus according to any preceding claim further comprising:
(d) one or more additional porous membranes positioned within said at least one first receptacle such that pore size of the porous membrane nearer the fluid inlet is greater than the pore size of the porous membrane nearer the fluid outlet.
15. An apparatus according to any preceding claim further comprising:
(e) a means for drawing liquid from the at least one first receptacle through the fluid outlet.
16. An apparatus according to claim 15 wherein the drawing means is a vacuum pump.
17. An apparatus according to any preceding claim further comprising:
(f) means for detecting binding of an analyte to the binding agent.
18. An apparatus according to claim 17 wherein the detecting means is selected from fluorimetric, chemiluminescent, radiochemical, physical and colourimetric means.
19. An apparatus according to claim 17 or 18 wherein the detecting means comprises a photometric device capable of detecting and/or quantifying light emitted from porous membrane (b).
20. An apparatus according to claim 19 wherein the photometric device is a luminometer.
21. An apparatus according to any one of claims 17-20 wherein the detecting means comprises imaging means.
22. An apparatus for use in an assay comprising a plurality of first receptacles in the form of a multi-well plate, characterised in that each receptacle comprises a plurality of immobilised analyte-specific binding agents.
23. An apparatus according to claim 22 wherein the plurality of analyte-specific binding agents is immobilised in the form of an array.
24. An apparatus according to claim 23 wherein the array is a micro-array.
25. An apparatus according to any one of claims 22-24 wherein the plurality of analyte-specific binding agents comprises at least one control probe.
26. An apparatus according to any one of claims 22-25 wherein said analyte-specific binding agents are polynucleotide probes.
27. An apparatus according to claim 26 wherein said analyte-specific binding agents are Expressed Sequence Tags (ESTs).
28. An apparatus according to any one of claims 22-25 wherein said analyte-specific binding agents are polypeptide binding agents.
29. An apparatus according to claim 28 wherein said analyte-specific binding agents are antibodies or a fragments thereof.
30. An apparatus according to any one of claims 22-29 wherein said analyte-specific binding agents are directly or indirectly immobilised onto a surface of said first receptacles.
31. An apparatus according to claim 30 wherein said surface is a fluid outlet.
32. An apparatus according to claim 31 wherein said surface is a porous membrane.
33. An apparatus according to claim 32 wherein said analyte-specific binding agent is immobilised onto the underside of said porous membrane.
34. An apparatus according to claim 32 or 33 wherein each first receptacle is positioned at least in part in a second receptacle which is capable of collecting liquid having passed through the fluid outlet of the first receptacle such that its porous membrane may be submerged in said liquid.
35. An apparatus according to any one of claims 32-34 comprising one or more further porous membranes positioned within at least one of said first receptacles such that membrane pore size gradually decreases towards the fluid outlet.
36. An apparatus according to any one of claims 30-35 further comprising a means for drawing liquid from the first receptacles through the fluid outlet.
37. An apparatus according to claim 36 wherein the drawing means is a vacuum pump.
38. An apparatus according to any one of claims 22-37 further comprising means for detecting binding of an analyte to the binding agent.
39. An apparatus according to claim 38 wherein the detecting means is selected from fluorimetric, chemiluminescent, radiochemical, physical and colourimetric means.
40. An apparatus according to claim 38 or 39 wherein the detecting means comprises a photometric device capable of detecting and/or quantifying light emitted from at least one of the plurality of first receptacles.
41. An apparatus according to claim 40 wherein the photometric device is a luminometer.
42. An apparatus according to any one of claims 38-41 wherein the detecting means comprises imaging means.
43. An apparatus according to any one of claims 21-42 in the form of a 96-well plate.
44. A method of detecting an analyte comprising:
(a) providing an apparatus according to any one of claims 1-42;
(b) loading a sample to be tested into the or at least one of the first receptacles;
and (c) detecting binding of the analyte to the analyte-specific binding agent.
45. A method according to claim 44 further comprising the step, between (b) and (c), of actively drawing liquid from the one or more first receptacles through the fluid outlet.
46. A method according to claim 45 wherein a vacuum pump is used to draw the liquid from the one or more receptacles.
47. A method according to any one of claims 44-46 wherein the sample comprises a detectable label.
48. A method according to claim 47 wherein the detectable label is selected from an enzyme, a fluorescent label and a radiolabel.
49. A method according to any one of claims 44-48 wherein step (c) comprises detecting binding of the analyte(s) to the one or more analyte-specific binding agents using any one of fluorimetric, chemiluminescent, radiochemical, physical and colourimetric means.
50. A method according to any one of claims 47-49 wherein step (c) comprises detecting binding of the analyte(s) to the one or more analyte-specific binding agents using a photometric device capable of detecting and/or quantifying light emissions.
51. A method according to claim SO wherein the photometric device is a luminometer.
52. A method according to any one of claims 47-51 wherein step (c) further or alternatively comprises individually detecting and/or quantifying light emissions from each spot using imaging means.
53. A method according to any one of claims 44-52 whereby, if the apparatus of (a) has more than one first receptacle, a different sample is loaded to each receptacle.
54. A method of detecting an analyte comprising:
(a) providing an apparatus comprising at least one first receptacle having a fluid inlet and a fluid outlet and having a porous membrane;

(b) immobilising at least one analyte-specific binding agent in a spot on the underside of said porous membrane;
(c) loading a sample to be tested into the at least one first receptacle; and (d) detecting binding of the analyte to the analyte-specific binding agent(s).
55. A method according to claim 54 wherein the porous membrane is positioned at the fluid outlet.
56. A method according to claim 54 or 55 wherein the apparatus comprises a plurality of first receptacles in the form of a multi-well plate.
57. A method according to claim 56 wherein the apparatus is in the form of a well plate.
58. A method according to any one of claims 54-57 wherein a plurality of analyte-specific binding agents is immobilised on said porous membrane in the form of an array.
59. A method according to claim 58 wherein said plurality of analyte-specific binding agents is immobilised in the form of a micro-array.
60. A method according to claim 58 or claim 59 wherein the array comprises at least one control probe.
61. A method according to any one of claims 54-60 wherein said at least one analyte-specific binding agent is a polynucleotide probe.
62. A method according to claim 61 wherein said at least one analyte-specific binding agent is an Expressed Sequence Tag (EST).
63. A method according to any one of claims 54-60 wherein said at least one analyte-specific binding agent is a polypeptide binding agent.
64. A method according to claim 63 wherein said at least one analyte-specific binding agent is an antibody or a fragment thereof.
65. A method according to any one of claims 54-64 wherein said at least one analyte-specific binding agent is immobilised either directly or indirectly on said porous membrane.
66. A method according to any one of claims 54-65 further comprising the step, between (c) and (d), of actively drawing liquid from the at least one first receptacle through the fluid outlet.
67. A method according to claim 66 wherein a vacuum pump is used to draw the liquid from the at least one first receptacle.
68. A method according to any one of claims 54-67 wherein the or each first receptacle of the apparatus is positioned at least in part in a second receptacle which is capable of collecting liquid drawn through the fluid outlet of the first receptacle such that its porous membrane is submerged in said liquid.
69. A method according to any one of claims 54-68 further comprising the step, between (c) and (d), of filtering the sample through one or more additional porous membranes positioned within said at least one first receptacle.
70. A method according to claim 69 wherein the one or more additional porous membranes are positioned such that membrane pore size gradually decreases from fluid inlet to fluid outlet.
71. A method according to any one of claims 54-70 wherein the sample comprises a detectable label.
72. A method according to claim 71 wherein the detectable label is selected from an enzyme, a fluorescent label and a radiolabel.
73. A method according to any one of claims 54-72 wherein step (d) comprises detecting binding of the analyte to the analyte-specific binding agent using any one of fluorimetric, chemiluminescent, radiochemical, physical and colourimetric means.
74. A method according to any one of claims 54-73 wherein step (d) comprises detecting binding of the analyte to the analyte-specific binding agent using a photometric device capable of detecting light emissions.
75. A method according to claim 74 wherein the photometric device is a luminometer.
76. A method according to any one of claims 54-73 wherein step (d) further or alternatively comprises individually detecting and/or quantifying light emitted from each spot using imaging means.
77. A method according to any one of claims 54-76 wherein, in step (c), a different sample is loaded to each first receptacle.
78. A method of detecting an analyte comprising:
providing an apparatus comprising a plurality of first receptacles in the form of a multi-well plate:
forming a plurality of spots of immobilised analyte-specific binding agent in at least one of said first receptacles;
loading a sample to be tested into said first receptacles; and detecting binding of the analyte to the analyte-specific binding agent.
79. A method according to claim 78 wherein the apparatus is in the form of a well plate.
80. A method according to claim 78 or claim 79 wherein the plurality of analyte-specific binding agents is immobilised in the form of an array.
81. A method according to claim 80 wherein the array is a micro-array.
82. A method according to any one of claims 78-81 wherein the plurality of analyte-specific binding agents comprises at least one control probe.
83. A method according to any one of claims 78-82 wherein said analyte-specific binding agents are polynucleotide probes.
84. A method according to claim 83 wherein said analyte-specific binding agents are Expressed Sequence Tags (ESTs).
85. A method according to any one of claims 78-82 wherein said analyte-specific binding agents are polypeptide binding agents.
86. A method according to claim 85 wherein said analyte-specific binding agents are antibodies or fragments thereof.
87. A method according to any one of claims 78-86 wherein said analyte-specific binding agents are directly or indirectly immobilised onto a surface of said first receptacles.
88. A method according to claim 87 wherein said surface is a fluid outlet.
89. A method according to claim 88 wherein said surface is a porous membrane.
90. A method according to claim 89 wherein said analyte-specific binding agent is immobilised onto the underside of said porous membrane.
91. A method according to claim 89 or 90 wherein each first receptacle is positioned at least in part in a second receptacle which is capable of collecting liquid having passed through the fluid outlet of the first receptacle such that its porous membrane may be submerged in said liquid.
92. A method according to any one of claims 88-91 further comprising the step, between (c) and (d), of filtering the sample through one or more additional porous membranes positioned within at least one of said first receptacles.
93. A method according to claim 92 wherein the one or more additional porous membranes are positioned such that membrane pore size gradually decreases towards the fluid outlet.
94. A method according to any one of claims 88-93 further comprising the step, between (c) and (d), of actively drawing liquid from the first receptacles through the fluid outlet.
95. A method according to claim 94 wherein a vacuum pump is used to draw the liquid from the receptacles.
96. A method according to any one of claims 78-95 wherein the sample comprises a detectable label.
97. A method according to claim 96 wherein the detectable label is selected from an enzyme, a fluorescent label and a radiolabel.
98. A method according to any one of claims 78-97 wherein step (d) comprises detecting binding of the analyte to the analyte-specific binding agent using any one of fluorimetric, chemiluminescent, radiochemical, physical and colourimetric means.
99. A method according to any one of claims 78-98 wherein step (d) comprises detecting binding of the analyte to the analyte-specific binding agent using a photometric device capable of detecting and/or quantifying light emissions.
100. A method according to claim 99 wherein the photometric device is a luminometer.
101. A method according to any one of claims 78-100 wherein step (d) further or alternatively comprises individually detecting and/or quantifying light emitted from each spot using imaging means.
102. A method according to any one of claims 78-101 wherein, in step (c), a different sample is loaded to each first receptacle.
103. A porous membrane for use in an apparatus according to any one of claims 1 to 43 or for use in a method according to any one of claims 44 to 102 wherein the porous membrane has at least one analyte-specific binding agent immobilised on one side thereof.
104. A kit for use in an apparatus according to any one of claims 1 to 43 or for use in a method according to any one of claims 44 to 102, wherein the kit comprises a porous membrane having at least one analyte-specific binding agent immobilised on one side thereof and at least one reagent for detecting binding of an analyte to the at least one analyte-specific binding agent.
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CA2524928C (en) 2012-01-17
US7700373B2 (en) 2010-04-20
US20070184494A1 (en) 2007-08-09
GB0311902D0 (en) 2003-06-25
WO2004103939A2 (en) 2004-12-02
GB0411381D0 (en) 2004-06-23
EP1628768B1 (en) 2012-04-18
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EP1628768A2 (en) 2006-03-01
JP4536727B2 (en) 2010-09-01
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JP2007505326A (en) 2007-03-08
ATE553845T1 (en) 2012-05-15

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